Primary Oligodendrocyte Culture and Isolation Service

Oligodendrocytes are specialized glial cells in the nervous system that support neuronal signaling through myelination. Myelination refers to the development of a protective fatty sheath called myelin that wraps around the axons of neurons. Myelin significantly improves the speed and efficiency of electrical signal transmission within the nervous system, making it essential for normal brain function and overall neurological health.

Creative Biolabs offers reliable primary oligodendrocyte culture models, essential for studying myelination and a range of demyelinating diseases (e.g., multiple sclerosis) and neurodegenerative diseases (e.g., Alzheimer's disease).

Acquisition of Oligodendrocyte Lineage Cells

Compared to neurons and microglia, obtaining high-purity oligodendrocyte precursor cells (OPCs) is more challenging. Recently, Creative Biolabs successfully achieved more than 95% purity of OPCs from the brains of postnatal rodents using the E3 method developed by Kim H et al¹.

In comparison to the shaking method pioneered by McCarthy & de Vellis, the E3 method offers higher cell yield and shorter time required to obtain pure OPCs, while significantly reducing potential cell damage that can occur from overnight shaking. Additionally, when compared to methods like immunopanning and magnetic-associated cell sorting (MACS), the E3 method demonstrates superior cost-effectiveness.

Fig. 1 Workflow for the E3 method for primary oligodendrocyte culture.^{1, 2}

Isolated OPCs could proliferate and differentiate into mature oligodendrocytes in 14 days, which are positive for MBP.

Fig. 2 Representative images of mouse oligodendrocytes cultured using the E3 method.^{1, 3}

Co-culturing passaged OPCs with neuron cultures, we observed the efficient formation of myelin sheaths.

Fig. 3 Myelination capacity of cultured oligodendrocytes in vitro.^{1, 3}

Applications of Primary Oligodendrocytes in Research

• Myelination, Demyelination and Remyelination Studies

Primary oligodendrocytes are crucial for studying myelination, demyelination, and remyelination processes. They serve as a model to investigate the mechanisms of myelin formation and the impact of injuries on myelin integrity, aiding in the understanding of oligodendrocyte biology and axonal protection.

• Neurodegenerative Diseases

Oligodendrocytes are crucial in conditions like multiple sclerosis, amyotrophic lateral sclerosis, and leukodystrophies, as these diseases involve damage to myelin or oligodendrocytes, leading to impaired nerve function. Studying primary oligodendrocytes can help elucidate the cellular mechanisms involved in these diseases and contribute to developing therapeutic strategies.

• Axon-Oligodendrocyte Interactions

Understanding the interactions between oligodendrocytes and axons is crucial for grasping how myelination affects neuronal health and function. Researchers can investigate how axonal signals affect oligodendrocyte survival and myelin formation.

• Cellular Models for Drug Testing

Primary oligodendrocytes are used as cellular models in pharmacological studies to test potential treatments for demyelinating diseases, assessing both the protective effects on oligodendrocytes and the restoration of myelination.

Common Assays Utilize Primary Oligodendrocytes

Myelination Assay

Purpose:

• Aim to identify compounds that promote myelination, an essential process for the development, maintenance, and repair of the nervous system.

Methods:

• Co-culture systems where primary oligodendrocytes are cultured with primary brain neurons, DRG neurons, retinal ganglion cells, or Schwann cells.
• Use of specific markers such as myelin basic protein (MBP) or proteolipid protein (PLP) to assess myelin formation via immunofluorescence staining or Western blotting
• Ultrastructural analysis of the number of myelinated axons and thickness of myelin sheaths was performed using electron microscopy (EM)
• Measurement of electrical properties of axons before and after myelination using techniques like electrophysiology.

OPC Differentiation Assay

Purpose:

• To evaluate the differentiation process of OPCs into mature oligodendrocytes. This is a critical phase in both development and the regeneration of the myelin sheath.

Methods:

• Culturing OPCs in media containing differentiation factors (e.g., T3 or interested compounds) and assessing morphological changes.
• Quantifying the expression of differentiation markers such as O4, NG2, PDGFRα and MBP, using flow cytometry or immunofluorescence.

OPC Migration Assay

Purpose:

• To evaluate the ability of oligodendrocytes to migrate toward different stimuli, which is vital for remyelination after injury or in diseases such as multiple sclerosis.

Methods:

• Using a Boyden chamber or Transwell to quantify migration in response to specific chemokines or growth factors.
• Assessing cellular motility with time-lapse imaging and analyzing the tracks using software for quantification.

References

1. Kim, Hanki et al. "A primary culture method for the easy, efficient, and effective acquisition of oligodendrocyte lineage cells from neonatal rodent brains." Heliyon. 2024;10(8):e29359.
2. Distributed under Open Access license CC BY 4.0. The original image was modified.
3. Distributed under Open Access license CC BY 4.0 without modification.